WO2017211016A1 - 一种鉴定有机葡萄酒的方法 - Google Patents

一种鉴定有机葡萄酒的方法 Download PDF

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WO2017211016A1
WO2017211016A1 PCT/CN2016/098134 CN2016098134W WO2017211016A1 WO 2017211016 A1 WO2017211016 A1 WO 2017211016A1 CN 2016098134 W CN2016098134 W CN 2016098134W WO 2017211016 A1 WO2017211016 A1 WO 2017211016A1
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wine
organic
flow rate
nitrogen
preset
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吴浩
靳保辉
陈波
金晓蕾
张建莹
颜治
谢丽琪
蔡屹
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深圳出入境检验检疫局食品检验检疫技术中心
深圳市检验检疫科学研究院
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Publication of WO2017211016A1 publication Critical patent/WO2017211016A1/zh

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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/02—Food
    • G01N33/14—Beverages
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/02—Food
    • G01N33/14—Beverages
    • G01N33/146—Beverages containing alcohol

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  • the invention relates to the field of product detection, and in particular to a method for identifying organic wine.
  • Organic wines are wines that receive the “Organic Grape Brewing” certification mark. It emphasizes the organicity of the raw materials of the wine, that is, the naturalness. Grapes come from organic vineyards or organically grown without chemical fertilizers and pesticides. Organic vineyards use natural materials for fertilizers for three consecutive years (such as seaweed, livestock manure and plant compost) and are harvested by hand. During the brewing process, special attention is paid to the use, filtration and clarification of natural yeast to improve the quality of organic wines. Organic wines do not contain chemical additives, and they cannot be used in plastic bottles or polyethylene.
  • the present invention provides a method of identifying an organic wine, and in particular, a method for identifying an organic wine using a nitrogen isotope.
  • a method of identifying an organic wine according to the present invention comprises the steps of:
  • the carrier gas flow rate is a preset flow rate
  • the oxygen flow rate is to a preset flow rate
  • the oxygen is passed for a predetermined time
  • step 1) freeze-dried sample is placed in an elemental analyzer, and the nitrogen stable isotope ratio is analyzed according to the analytical standard method;
  • step 6) Determine whether the wine is an organic wine based on the data analyzed in step 5) and the preset threshold.
  • step 3 the temperature of the elemental analyzer reaction tube is raised to 960 degrees Celsius, the carrier gas flow rate is 110 mL/min, the oxygen flow rate is 175 mL/min, and the oxygen-passing time is set to 3 seconds.
  • step 5) the method further comprises the steps of: establishing a stable range of nitrogen isotope ratios of the organic wine, and setting the organic wine ⁇ 15 N>3.5 ⁇ to determine the organic wine.
  • 1 is a comparison data of organic wines and non-organic wines obtained by the method for identifying organic wines provided by an embodiment of the present invention.
  • the principle of the present invention is that the chemical fertilizer mainly utilizes N 2 synthetic nitrogen fertilizer in the atmosphere, so that the stable nitrogen isotope of the chemical fertilizer is close to the atmosphere, between -2 ⁇ and 2 ⁇ , and animal fat such as poultry manure
  • the average nitrogen stable isotope ratio is 8 ⁇ , up to 36 ⁇ .
  • the nitrogen isotope of the grapes produced by the fertilizers is significantly lower than that of the organic fertilizer sources.
  • the invention removes moisture and volatile alcohols (ethanol, butanol, etc.) in the wine by freezing or heating, collects the remaining solid samples, and then measures the sample by elemental analysis-stable isotope mass spectrometry.
  • the stable isotope ratio of nitrogen in the product Combined with the true organic wine nitrogen stable isotope distribution threshold, determine whether the sample is organic wine.
  • Experimental procedure of the present invention (1) 5 mL of a wine sample is taken and freeze-dried in a drying dish. (2) Take about 2mg of the sample after drying, put it in a tin cup, and wrap it tightly. (3) Open the stable isotope ratio mass spectrometer host and adjust the reference gas balance so that the 10 sets of N 2 reference gas nitrogen stable isotope ratio variation is less than 0.06 ⁇ . (4) Open the elemental analyzer combined with the stable isotope ratio mass spectrometer, raise the temperature of the elemental analyzer reaction tube to 960 degrees Celsius, the carrier gas flow rate of 110 mL/min, the oxygen flow rate of 175 mL/min, and set the oxygenation time to 3 seconds.
  • the isotope standard IAEA-600 provided by IAEA is used as a standard, and a standard sample of about 0.1 mg is wrapped in a tin cup to analyze the stable isotope ratio of the standard nitrogen until the average of the three measured values.
  • the difference from the standard value is ⁇ 15 N ⁇ 0.5 ⁇ .
  • the sample in the step (2) is placed in an elemental analyzer, and the nitrogen stable isotope ratio is analyzed according to the analytical standard method.
  • the nitrogen isotope was used as a judgement for judging whether it was an organic wine; a wine solid sample was prepared by freeze drying and heating, and the nitrogen stable isotope ratio was analyzed by an elemental analysis-stabilized nitrogen-nose ratio mass spectrometer.
  • the true organic wine has a nitrogen stable isotope ratio of ⁇ 15 N>3.5 ⁇ .
  • the standard IAEA-600 (caffeine) supplied by the International Atomic Energy Agency was selected, and 0.1 mg was weighed in a tin cup.
  • the results of 10 analyses are shown in Table 1 (unit: ⁇ ). It can be seen from Table 1 that the mean value is 0.94 ⁇ , which is 0.06 ⁇ from the true value of the standard product, and the standard deviation (SD) is 0.11 ⁇ , which is better than the instrument uncertainty of 0.2 ⁇ , which indicates that the instrument analysis error is very small and meets the analytical requirements.
  • the organic wine samples are collected from the 100,000-mu organic grape industrial park in Ningxia Delong, located in the Helan Mountain and Vineyards.
  • the vineyard has obtained the China Organic Certification (certification number: CHC11010178R1M-1) and the US USDA organic certification (certification number: 7327CN100 (NOP)).
  • CHC11010178R1M-1 China Organic Certification
  • US USDA organic certification certification number: 7327CN100 (NOP)
  • the whole process is organic, no pesticides and fertilizers are used in the planting process, and livestock manure is used as organic fertilizer (mainly beef and manure).
  • Non-organic wines are collected from Helan Shandong ⁇ non-organic winery, specific Sample information is shown in Table 1.
  • Non-organic wines are made from Cabernet Sauvignon, which is dominated by Cabernet Sauvignon, Merlot, Syrah, Cabernet Gernischt and Riesling. All grape varieties belong to the Vitis vinifera variety.
  • the minimum stable isotope ratio of organically grown organic wines is 3.5 ⁇ , and the average value reaches 4.23 ⁇ . Therefore, the organic wine nitrogen stable isotope ratio threshold is set at a minimum value.
  • the description of the term "one embodiment” or the like means that a specific feature, structure, material or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention.
  • the schematic representation of the above terms does not necessarily refer to the same embodiment or example.
  • the particular features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments or examples.

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Abstract

一种鉴定有机葡萄酒的方法，包括：1)将待测葡萄酒样品冷冻干燥；2)调节参考气平衡，使多组N 2参考气氮稳定同位素比值变异小于0.06‰；3)将元素分析仪反应管温度调节至预设温度，载气流速度为预设流速，氧气流速至预设流速，通氧预定时间；4)分析标准品氮稳定同位素比值，直到至少3次测定值的均值与标准值差异δ 15N<0.5‰；5)将步骤1)冷冻干燥后的样品放入元素分析仪中，按照分析标准品方法分析氮稳定同位素比值；6)并根据步骤5)中分析的数据和预设的阈值，判断葡萄酒是否为有机葡萄酒。所述方法仅需分析一个指标即能判断葡萄酒是否为有机肥料种植生产的，前处理简单，分析过程能在10分钟内完成。

Description

一种鉴定有机葡萄酒的方法 技术领域
本发明涉及品检测领域，具体地涉及一种鉴定有机葡萄酒的方法。
背景技术
本部分中的陈述仅仅提供了与本发明公开的内容有关的背景信息，且可能不构成现有技术。
有机葡萄酒是指获得“有机葡萄酿造”认证商标的葡萄酒。其特别强调酿酒原料的有机性，即纯天然性。葡萄来自有机葡萄园，或采取有机种植法，一概不用化学肥料和农药。有机葡萄园连续3年采用天然的物质作肥料(如海藻、牲口粪便和植物混合肥料)，并以人工采收。在酿制过程中，特别注意天然酵母的使用、过滤和澄清方法，以提高有机葡萄酒的质量。有机葡萄酒中不含化学添加剂，更不能以塑料瓶、聚乙烯等来盛酒。
然而，现有技术并没有相应的技术去鉴定葡萄酒为有机葡萄酒或者无机葡萄酒。
因此，现有技术有待改进和发展。
发明内容
为了解决现有技术中的上述问题，本发明提供了一种鉴定有机葡萄酒的方法，具体地提供了一种使用氮同位素鉴定有机葡萄酒的方法。
根据本发明提供的一种鉴定有机葡萄酒的方法，其包括以下步骤：
1)将待测葡萄酒样品冷冻干燥；
2)调节参考气平衡，使多组N2参考气氮稳定同位素比值变异小于0.06‰；
3)将元素分析仪反应管温度调节至预设温度，载气流速度为预设流速，氧气流速至预设流速，通氧预定时间；
4)分析标准品氮稳定同位素比值，直到至少3次测定值的均值与标准值差异δ15N<0.5‰；
5)将步骤1)冷冻干燥后的样品放入元素分析仪中，按照分析标准品方法分析氮稳定同位素比值；
6)并根据步骤5)中分析的数据和预设的阈值，判断葡萄酒是否为有机葡萄酒。
进一步地，步骤3)中元素分析仪反应管温度提升至960摄氏度，载气流110mL/min，氧气流速175mL/min，设置通氧时间为3秒。
进一步地，在步骤5)和步骤6)之间还包括如下步骤：建立有机葡萄酒氮稳定同位素比值范围，设定有机葡萄酒δ15N>3.5‰则判断为有机葡萄酒。
根据本发明提供的一种鉴定有机葡萄酒的方法，能够带来至少以下有益效果：
仅需分析一个指标即能判断葡萄酒是否为有机肥料种植生产的有机葡萄，前处理简单，分析过程能在10分钟内完成。
附图说明
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案，下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图是本发明的一些实施方式，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为采用本发明实施例提供的鉴定有机葡萄酒的方法而获得的有机葡萄酒与非有机葡萄酒的比较数据。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚，下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
参照图1，本发明的原理为：化肥主要是通过利用大气中的N2合成氮肥，因此化肥氮稳定同位素与大气接近，介于-2‰～2‰之间，而动物肥如家禽粪肥的氮稳定同位素比值平均值达到8‰，最高可达36‰。因此导致化肥种植的葡萄酿的葡萄酒氮同位素显著低于有机肥来源的葡萄酒。通过分析葡萄酒中氮稳定同位素比值即可判断该葡萄酒是否为有机葡萄酒。
本发明通过冷冻或加热将葡萄酒中水分和挥发性醇类(乙醇、丁醇等)除去，收集剩余的固体样品，再通过元素分析－稳定同位素质谱仪测定样 品中的氮稳定同位素比率。结合真实有机葡萄酒氮稳定同位素分布阈值，判断样品是否为有机葡萄酒。
本发明实验步骤：(1)取葡萄酒样品5mL，于干燥皿中冷冻干燥。(2)取干燥后样品2mg左右，置于锡杯中，包裹严密待测。(3)打开稳定同位素比率质谱仪主机，调节参考气平衡，使10组N2参考气氮稳定同位素比值变异小于0.06‰。(4)打开与稳定同位素比率质谱仪联用的元素分析仪，将元素分析仪反应管温度提升至960摄氏度，载气流110mL/min，氧气流速175mL/min，设置通氧时间为3秒。(5)仪器达到稳定状态后，以IAEA提供的同位素标准品IAEA-600为标准品，包裹0.1mg左右的标准样品于锡杯中，分析标准品氮稳定同位素比值，直到3次测定值的均值与标准值差异δ15N<0.5‰。(6)将步骤(2)中的样品放入元素分析仪中，按照分析标准品方法分析氮稳定同位素比值。(7)建立有机葡萄酒氮稳定同位素比值范围，本发明根据已有数据库数据，设定有机葡萄酒δ15N>3.5‰则判断为有机葡萄酒。(8)根据步骤(6)中分析的数据和步骤(7)中的阈值，判断葡萄酒是否为有机葡萄酒。
仅需分析一个指标即能判断葡萄酒是否为有机肥料种植生产的有机葡萄，前处理简单，分析过程能在10分钟内完成。
使用氮同位素作为判断是否为有机葡萄酒的判定指标；使用冷冻干燥和加热方法制备葡萄酒固体样，并用元素分析-稳定氮同素比率质谱仪分析氮稳定同位素比值。真实有机葡萄酒的氮稳定同位素比值为δ15N>3.5‰。
数据1、氮稳定同位素准确性分析
选取国际原子能机构提供的标准品IAEA-600(咖啡因)，称取0.1mg用锡杯包裹，10次分析结果见表1(单位为‰)。由表1可知，均值为0.94‰，与标准品真值相差0.06‰，标准偏差(SD)为0.11‰，优于仪器不确定度0.2‰，表明仪器分析误差极小，满足分析要求。
表1 IAEA-600标准品10次分析数据(标准值δ15N＝1.0)
Figure PCTCN2016098134-appb-000001
数据2、稳定性分析
随机选取5个葡萄酒样品，每个样品测定5次氮稳定同位素比值。由表2可知，5个样品分析的标准偏差均优于0.5‰，满足分析要求。
表2样品分析稳定性数据
  1 2 3 4 5 平均值(‰) SD(‰)
样品1 1.22 1.12 1.62 1.53 1.15 1.33 0.23
样品2 3.05 3.12 3.23 3.35 3.07 3.16 0.12
样品3 5.31 5.68 5.19 5.44 5.49 5.42 0.19
样品4 3.53 3.78 3.56 3.37 3.79 3.61 0.18
样品5 2.87 2.50 2.69 2.25 2.01 2.46 0.34
数据3、有机认证葡萄园有机葡萄酒数据
有机葡萄酒样品采自宁夏德龙十万亩有机葡萄产业园，位于贺兰山东麓葡萄园产区。该葡萄园获得中国有机认证(认证号：CHC11010178R1M-1)和美国USDA有机认证(认证号：7327CN100(NOP))。管理过程中实现全程有机，种植过程中不使用农药和化肥，以家畜粪肥作为有机肥料(主要为牛羊粪肥)。非有机葡萄酒采自贺兰山东麓非有机种植葡萄酒厂，具体 样品信息见表1。非有机葡萄酒使用赤霞珠(Cabernet Sauvignon)酿造，有机葡萄品种主要有赤霞珠(Cabernet Sauvignon)、梅洛(Merlot)、西拉(Syrah)、蛇龙珠(Cabernet Gernischt)和雷司令(Riesling)，所有的葡萄品种均属于欧亚葡萄品种(Vitis vinifera)。
表3样品信息
Figure PCTCN2016098134-appb-000002
由表4(单位为‰)可知，有机种植的有机葡萄酒氮稳定同位素比值最小值为3.5‰，且平均值达到4.23‰。因此以最小值设置有机葡萄酒氮稳定同位素比值阈值。
表4有机葡萄园葡萄酒数据
Figure PCTCN2016098134-appb-000003
Figure PCTCN2016098134-appb-000004
数据4、有机与非有机葡萄酒数据比较
非有机种植与有机种植生产的葡萄酒氮稳定同位素比值差异见图1，非有机种植生产的葡萄酒和有机种植生产的葡萄酒的δ15N分别为2.20±0.40‰和4.36±0.58‰。经过独立样品t-test统计分析，有机葡萄酒氮稳定同位素显著高于非有机种植葡萄酒(p<0.01)。
在本说明书的描述中，术语“一个实施例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或特点包含于本发明的至少一个实施例或示例中。在本说明书中，对上述术语的示意性表述不一定指的是相同的实施例或实例。而且，描述的具体特征、结构、材料或特点可以在任何的一个或多个实施例或示例中以合适的方式结合。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (3)

  1. 一种鉴定有机葡萄酒的方法，其特征在于，包括以下步骤：
    1)将待测葡萄酒样品冷冻干燥；
    2)调节参考气平衡，使多组N2参考气氮稳定同位素比值变异小于0.06‰；
    3)将元素分析仪反应管温度调节至预设温度，载气流速度为预设流速，氧气流速至预设流速，通氧预定时间；
    4)分析标准品氮稳定同位素比值，直到至少3次测定值的均值与标准值差异δ15N<0.5‰；
    5)将步骤1)冷冻干燥后的样品放入元素分析仪中，按照分析标准品方法分析氮稳定同位素比值；
    6)并根据步骤5)中分析的数据和预设的阈值，判断葡萄酒是否为有机葡萄酒。
  2. 根据权利要求1所述的鉴定有机葡萄酒的方法，其特征在于，步骤3)中元素分析仪反应管温度提升至960摄氏度，载气流110mL/min，氧气流速175mL/min，设置通氧时间为3秒。
  3. 根据权利要求1所述的鉴定有机葡萄酒的方法，其特征在于，在步骤5)和步骤6)之间还包括如下步骤：建立有机葡萄酒氮稳定同位素比值范围，设定有机葡萄酒δ15N>3.5‰，则判断为有机葡萄酒。
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